Auto cascade refrigeration (ACR) cycle with phase separators is widely used in the cryogenic system. The composition of mixed refrigerant has a great effect on the performance of the system. Based on the assumption of infinite volume of phase separator, ACR system with one phase separator is simulated in this paper. The variation of refrigerant composition under different valves opening is obtained. A related experimental system is set up to verify the variation. The result shows that when the valve opening connected to the evaporator increases or the valve opening under the phase separator decreases, the low-boiling component concentration of the working mixture passing through the compressor and condenser increases, while the high-boiling component concentration decreases. Furthermore, the variations of condensation pressure and evaporation pressure under different valves opening are also observed. This paper is helpful to deepen the understanding of ACR system. (C) 2014 The Chemical Industry and Engineering Society of China, and Chemical Industry Press. All rights reserved.